| 货号 | 7132C |
| 描述 | The PathScan® Total Akt1 Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total Akt1 protein with a chemiluminescent readout. Chemiluminescence ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA which is offered in low volume microplates, shows increased signal and sensitivity while using smaller sample size. An Akt rabbit antibody has been coated on the microwells. After incubation with cell lysates, the Akt protein is captured by the coated antibody. Following extensive washing, an Akt1 mouse antibody is added to detect the captured total Akt1 protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of total Akt1 protein. |
| 反应种属 | Human/Mouse |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Total Akt1 Chemiluminescent Sandwich ELISA Kit #7132 detects endogenous levels of total Akt1 protein in human and mouse cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species. |
| 供应商 | CST |
| 背景 | Akt, also referred to as PKB or Rac, plays a critical role in controlling survival and apoptosis (1-3). This protein kinase is activated by insulin and various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (2,3). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (4) and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2 responsible for phosphorylation of Akt at Ser473 has been identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 (5,6). Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including Bad (7), forkhead transcription factors (8), c-Raf (9), and caspase-9. PTEN phosphatase is a major negative regulator of the PI3 kinase/Akt signaling pathway (10). LY294002 is a specific PI3 kinase inhibitor (11). Another essential Akt function is the regulation of glycogen synthesis through phosphorylation and inactivation of GSK-3α and β (12,13). Akt may also play a role in insulin stimulation of glucose transport (12). In addition to its role in survival and glycogen synthesis, Akt is involved in cell cycle regulation by preventing GSK-3β-mediated phosphorylation and degradation of cyclin D1 (14) and by negatively regulating the cyclin dependent kinase inhibitors p27 Kip (15) and p21 Waf1/CIP1 (16). Akt also plays a critical role in cell growth by directly phosphorylating mTOR in a rapamycin-sensitive complex containing raptor (17). More importantly, Akt phosphorylates and inactivates tuberin (TSC2), an inhibitor of mTOR within the mTOR-raptor complex (18,19). |
| 存放说明 | 4C |
| 参考文献 | Franke, T.F. et al. (1997) Cell 88, 435-7. Burgering, B.M. and Coffer, P.J. (1995) Nature 376, 599-602. Franke, T.F. et al. (1995) Cell 81, 727-36. Alessi, D.R. et al. (1996) EMBO J 15, 6541-51. Sarbassov, D.D. et al. (2005) Science 307, 1098-101. Jacinto, E. et al. (2006) Cell 127, 125-37. Cardone, M.H. et al. (1998) Science 282, 1318-21. Brunet, A. et al. (1999) Cell 96, 857-68. Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-4. Cantley, L.C. and Neel, B.G. (1999) Proc Natl Acad Sci USA 96, 4240-5. Vlahos, C.J. et al. (1994) J Biol Chem 269, 5241-8. Hajduch, E. et al. (2001) FEBS Lett 492, 199-203. Cross, D.A. et al. (1995) Nature 378, 785-9. Diehl, J.A. et al. (1998) Genes Dev 12, 3499-511. Gesbert, F. et al. (2000) J Biol Chem 275, 39223-30. Zhou, B.P. et al. (2001) Nat Cell Biol 3, 245-52. Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31. Inoki, K. et al. (2002) Nat Cell Biol 4, 648-57. Manning, B.D. et al. (2002) Mol Cell 10, 151-62. |
Relationship between protein concentration of lysates from untreated and PDGF-treated NIH/3T3 cells and immediate light generation with chemiluminescent substrate is shown. Cells (80% confluence) were treated with PDGF #9909 (50 ng/ml) and lysed after incubation at 37ºC for 20 minutes. Graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.未处理和PDGF处理组NIH/3T3细胞提取物中蛋白浓度与化学发光底物生成的即发光的线性关系图。NIH/3T3细胞（80%）采用PDGF F #990(50 ng/ml)在37度处理20分钟，然后裂解。插图相对应的阴影部分显示在低蛋白浓度范围的高灵敏度和线性反应。 |
